448 JOURNAL OF COSMETIC SCIENCE
This article highlights five new commercially available technologies that meet sustainable
solution requirements while also providing exceptional performance. These technologies
include three novel active ingredients such as the Zenerity™ (Lubrizol, Brecksville, OH,
USA) biotech ingredients (INCI: glycerin, water (aqua), and bacillus ferment THW
biotech ingredient), Uplevity™ (Lipotec USA, Lewisville, TX, USA) lift peptide (INCI:
tetrapeptide-1), and Stevisse™ advanced botanical ingredients (INCI: glycerin, water
(aqua), Stevia rebaudiana leaf/stem extract), as well as two novel rheology modifiers in the
Carbopol® Fusion (Lubrizol, Brecksville, OH, USA) S-20 polymer (INCI: starch acetate/
adipate (SAA) and citric acid)and Sensomer™ (Lubrizol, Brecksville, OH, USA) tara
polymer (INCI: Caesalpinia spinosa gum tara). The resulting sustainability and performance
of these products will be discussed first for the active ingredients and then for the rheology
modifiers.
MATERIALS AND METHODS
PREPARATION OF BOTANICAL EXTRACTS:
1. Subcritical water extract of stevia leaves was prepared using an accelerated solvent
extractor (AE350, Dionex, Sunnyvale, CA, USA) under the following general conditions.
Approximately 5 g of the botanical raw material (stevia leaves) were loaded on to a
100 mL stainless steel cell along with a layer of diatomaceous earth (10 g, ASE prep DE,
Dionex). The mixture was then placed in the extractor, filled with water, and heated to
temperatures in the range of 150o-180oC under high pressure (up to 1.38 MPa) for 15
minutes to maintain subcritical water conditions. The process was repeated three times.
The combined extract was subsequently mixed with glycerin to make a 60% glycerin
solution (weight basis).
2. A hot water extract was prepared by a steeping process using 5g of the botanical raw
material and 100 mL of water. The mixture was heated and mixed at 45°C for 4 hours.
The water extract was filtered and diluted with glycerin to make a 60% glycerin solution.
The aqueous glycerin extractions were produced similarly to the hot water process but
with a 60% glycerin aqueous solution at the exact same temperature of 45°C.
ORGANIZATION FOR ECONOMIC COOPERATION AND DEVELOPMENT (OECD) 301
BIODEGRADABILITY TEST
This test guideline describes six methods that permit screening chemicals for ready
biodegradability in an aerobic aqueous medium. These methods include: the dissolved
organic content (DOC) die-away, the CO
2 evolution (modified Sturm test), the MITI (I)
(Ministry of International Trade and Industry, Japan), the closed bottle, the modified
OECD screening, and the manometric respirometry. A solution or suspension of the well
determined/described test substance in a mineral medium is inoculated and incubated
under aerobic conditions in the dark or in diffuse light. Running blanks with inoculum
but without a test substance in parallel helps to determine the endogenous activity of the
inoculum. A reference compound (aniline, sodium acetate, or sodium benzoate) is also run
in parallel to check the operation of the procedures. Normally, the test lasts for 28 days. At
least two flasks or vessels containing the test substance (plus the inoculum) and at least two
flasks or vessels containing only the inoculum should be used single vessels are sufficient

449 DELIVERING SUSTAINABLE SOLUTIONS TO IMPROVE WELLBEING
for the reference compound. In general, degradation is followed by determining parameters
like DOC, CO
2 production, and oxygen uptake. The pass levels for ready biodegradability
are 70% removal of DOC and 60% of theoretical oxygen demand or ThCO
2 production for
respirometric methods. These pass values must be reached in a 10-day window within the
28-day period of the test.1
INHIBITION OF HIGH MOBILITY GROUP PROTEIN B1 (HMGB1) STRESSORIN RELEASE IN HUMAN
EPIDERMAL KERATINOCYTES
Human epidermal keratinocytes from adults (HEKa cells) were seeded in 96-well plates in
culture medium. After 24 hours of incubation, the culture medium was removed, and a
metabolic stress was applied by adding 20 mM of glucose (Merck Life Science) and 0.1 mM
of palmitic acid (Merck Life Science) to the culture medium, simulating an unhealthy diet
in cell culture with or without treatment with a THW biotech ingredient concentrate
(without glycerin) at 100 µg/mL. Cells treated with culture medium, glucose, and palmitic
acid were used as a metabolic stress control. Cells treated with culture medium alone were
used as a control. After 24 hours of incubation, the supernatants were collected, and the
stressorin inhibition was measured using a stressorin marker (HMGB1) by ELISA Novus
Biologicals, Abingdon, United Kingdom) according to the manufacturer’s protocol. These
results were normalized with the total viable cell number for each condition calculated
by the PrestoBlueTM staining assay (Thermo Fisher Scientific, Waltham, MA, USA). The
obtained values were normalized compared to the control or metabolic stress control
as applicable and represent the mean of three independent experiments performed in
triplicates. The statistical analysis used was the unpaired student’s t test.
EVALUATION OF THE ANTIGLYCATION EFFECT
To assess the ability of the active ingredient to counteract the formation of advanced glycation
end products (AGEs), an in tubo glycation reaction was performed. Type I collagen (0.6 mg/
mL) was incubated with glucose at 0.4 M and with 100 µg/mL of THW biotech ingredient
concentrate at 60°C for 3 days. As a glycation control, only type I collagen and glucose at
the same concentrations were incubated, a condition in which glycation occurs. Glycation
levels were determined by measuring AGE-specific fluorescence, or 370(ex)/440(em) using
ClarioSTAR. Samples were tested in triplicates and assayed in four different replicates.
REDUCTION OF LIPOFUSCIN ACCUMULATION WITH AGE IN HUMAN EPIDERMAL KERATINOCYTES
Cellular garbage reduction was evaluated by analyzing lipofuscin accumulation (an
autofluorescent pigment commonly used as an indicator of waste accumulation) in
epidermal cells using autofluorescence. HEKa cells were seeded in 96-well cell carrier
black plates (PerkinElmer, Inc., Waltham, MA, USA) in culture medium. After 24 hours
of incubation, the culture medium was removed, and cells were incubated in culture
medium in combination with metabolic stress (20 mM glucose and 0.1 mM palmitic
acid), simulating an unhealthy diet in the cell culture with or without treatment with
the THW biotech ingredient concentrate at 100 µg/mL. At the same time, cells treated
with the culture medium alone were used as a control, and cells treated with culture